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Literature summary for 4.2.1.84 extracted from

  • Nagasawa, T.; Nanba, H.; Ryuno, K.; Takeuchi, K.; Yamada, H.
    Nitrile hydratase of Pseudomonas chlororaphis B23. Purification and characterization (1987), Eur. J. Biochem., 162, 691-698.
    View publication on PubMed
Search for more literature for 4.2.1.84

Application

Application Comment Organism
synthesis
-
 Pseudomonas chlororaphis

Crystallization (Commentary)

Crystallization (Comment) Organism
-
 Pseudomonas chlororaphis

General Stability

General Stability Organism
completely stabilized by 22 mM n-butyric acid Pseudomonas chlororaphis

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 1-5 mM Pseudomonas chlororaphis
2,2'-dipyridyl 1-5 mM Pseudomonas chlororaphis
8-hydroxyquinoline
-
 Pseudomonas chlororaphis
Ag+ 0.01-1 mM AgNO3 Pseudomonas chlororaphis
Cu2+ 1-5 mM CuSO4 Pseudomonas chlororaphis
cysteamine 1 mM Pseudomonas chlororaphis
Disodium 4,5-dihydroxy-m-benzene disulfonate 0.1-5 mM Pseudomonas chlororaphis
FeCl3 1-5 mM Pseudomonas chlororaphis
FeSO4 1-5 mM, weak inhibition Pseudomonas chlororaphis
H2O2 0.1 mM Pseudomonas chlororaphis
Hg2+ HgCl2, 0.01-1 mM Pseudomonas chlororaphis
hydroxylamine 1 mM Pseudomonas chlororaphis
NaN3 1 mM Pseudomonas chlororaphis
p-chloromercuribenzoate 0.1 mM Pseudomonas chlororaphis
PbCl2 1 mM Pseudomonas chlororaphis
Penicillamine 1 mM; L- and D-isomer Pseudomonas chlororaphis
phenylhydrazine 1 mM Pseudomonas chlororaphis
Semicarbazide 1 mM Pseudomonas chlororaphis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.03
-
 n-butyronitrile pH 7.0, 20ºC Pseudomonas chlororaphis
2.33
-
 n-valeronitrile pH 7.0, 20ºC Pseudomonas chlororaphis
3.8
-
 Methacrylonitrile
-
 Pseudomonas chlororaphis
22.2
-
 Chloroacetonitrile pH 7.0, 20ºC Pseudomonas chlororaphis
29.4
-
 Propionitrile pH 7.0, 20ºC Pseudomonas chlororaphis
34.6
-
 acrylonitrile pH 7.0, 20ºC Pseudomonas chlororaphis

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ contains approximately 4 mol iron/mol enzyme Pseudomonas chlororaphis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
25000
-
 4 * 25000, SDS-PAGE Pseudomonas chlororaphis
100000
-
 gel filtration Pseudomonas chlororaphis

Organism

Organism UniProt Comment Textmining
Pseudomonas chlororaphis
-
-
-
Pseudomonas chlororaphis B23
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, octyl and phenyl-Sepharose CL-4B Pseudomonas chlororaphis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Pseudomonas chlororaphis
1840
-
 crystallized enzyme Pseudomonas chlororaphis

Storage Stability

Storage Stability Organism
-20°C, 50% glycerol, 0.01 M HEPES/KOH buffer, pH 7.2, 22 mM n-butyric acid, stable for more than 2 months Pseudomonas chlororaphis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acrylonitrile + H2O
-
 Pseudomonas chlororaphis 2-propenoic acid amide i.e. acrylic acid amide ?
chloroacetonitrile + H2O
-
 Pseudomonas chlororaphis chloroacetamide
-
 ?
chloroacetonitrile + H2O
-
 Pseudomonas chlororaphis B23 chloroacetamide
-
 ?
isobutyronitrile + H2O
-
 Pseudomonas chlororaphis isobutyric acid amide
-
 ?
methacrylonitrile + H2O
-
 Pseudomonas chlororaphis methylacrylic acid amide
-
 ?
additional information specificity overview Pseudomonas chlororaphis ?
-
 ?
additional information aromatic nitriles are barely hydrated by the enzyme Pseudomonas chlororaphis ?
-
 ?
additional information formamide, acetamide, acrylamide, propionamide, n-butyramide, isobutyramide, n-valeramide, succinamide, benzamide, phenylacetamide and lactamide are no substrates Pseudomonas chlororaphis ?
-
 ?
additional information specificity overview Pseudomonas chlororaphis B23 ?
-
 ?
additional information aromatic nitriles are barely hydrated by the enzyme Pseudomonas chlororaphis B23 ?
-
 ?
additional information formamide, acetamide, acrylamide, propionamide, n-butyramide, isobutyramide, n-valeramide, succinamide, benzamide, phenylacetamide and lactamide are no substrates Pseudomonas chlororaphis B23 ?
-
 ?
n-butyronitrile + H2O
-
 Pseudomonas chlororaphis n-butyric acid amide
-
 ?
n-capronitrile + H2O
-
 Pseudomonas chlororaphis n-hexanoic acid amide
-
 ?
propionitrile + H2O
-
 Pseudomonas chlororaphis propionic acid amide
-
 ?
propionitrile + H2O
-
 Pseudomonas chlororaphis B23 propionic acid amide
-
 ?
valeronitrile + H2O
-
 Pseudomonas chlororaphis n-pentanoic acid amide
-
 ?

Subunits

Subunits Comment Organism
tetramer 4 * 25000, SDS-PAGE Pseudomonas chlororaphis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20
-
 with propionitrile and acrylonitrile as substrates Pseudomonas chlororaphis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 30 79% of activity maximum at 10°C, 13% of activity maximum at 30°C Pseudomonas chlororaphis
25 50 about 50% of activity maximum at 25°C, about 70% of activity maximum at 50°C Pseudomonas chlororaphis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
10
-
 10 min, 22 mM n-butyric acid, stable Pseudomonas chlororaphis
20
-
 10 min, 22 mM n-butyric acid, stable Pseudomonas chlororaphis
25
-
 10 min, 22 mM n-butyric acid, 12.5% loss of activity Pseudomonas chlororaphis
30
-
 10 min, 22 mM n-butyric acid, 25% loss of activity Pseudomonas chlororaphis
35
-
 1 hour, 10 mM methacrylonitrile stable up to this temperature Pseudomonas chlororaphis
35
-
 10 min, 22 mM n-butyric acid, 46.6% loss of activity Pseudomonas chlororaphis
40
-
 10 min, 22 mM n-butyric acid, 94.3% loss of activity Pseudomonas chlororaphis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
30.7
-
 Propionitrile
-
 Pseudomonas chlororaphis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Pseudomonas chlororaphis